Properties of an oviducal protein involved in amphibian oocyte fertilization.
The secretion produced at the most cephalic portion of the oviduct of the toad (pars recta in Bufo arenarum) is involved in fertilization. Although the present study indicates that a prerequisite for the fertilization of coelomic eggs is either their passage through the pars recta (PR) or their treatment with PR secretion fluid prior to insemination, the exact role of this secretion in the fertilization process is still not clearly understood. A protein acting upon the vitelline envelope (VE) of Bufo arenarum coelomic eggs has been purified from secretion fluid (pars recta protein, PRP). Some properties of both this protein and the secretion fluid are examined in an effort to understand their mechanism of action. It was shown that PRP partially dissolves the VE of coelomic oocytes in a way that resembles the action of proteolytic enzymes. PRP has also proved to be active on synthetic substrates of proteolytic enzymes such as p-Tosyl-L-arginine methyl ester-HCl (TAME), alpha-N-benzoyl-L-arginine ethyl ester-HCl (BAEE), and alpha-N-benzoyl-DL-arginine-p-nitroanilide HCl (BAPNA). PR enzyme is activated by calcium ions, shows a broad peak of maximum activity at pH 7.8, is stable at alkaline pH, and is inhibited by soybean trypsin inhibitor (SBTI) and N-alpha-p-tosyl-L-lysine chloromethyl ketone HCl (TLCK) but not by lima bean trypsin inhibitor (LBTI) or L-1-tosyl-amide-2-phenylethylchloro-methyl-ketone (TPCK).
D000818:Animals; D002022:Bufo arenarum; D002118:Calcium / Q000494:pharmacology; D005260:Female; D005306:Fertilization; D007700:Kinetics; D009865:Oocytes / Q000502:physiology*; D010057:Oviducts / Q000201:enzymology / Q000502:physiology*; D010063:Ovum / Q000502:physiology*; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000502:physiology*; D011480:Protease Inhibitors / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology
D011480:Protease Inhibitors; D014107:Tosyllysine Chloromethyl Ketone; D010447:Peptide Hydrolases; D002118:Calcium